Dengue is a vector-borne disease caused by dengue virus. According to the recent report of CDC that one-third population of the world are at high risk with Dengue fever. The prevalence of the dengue hemorrhagic fever was found more in tropical and sub-tropical regions of the world. Aedes mosquitoes was reported as the main cause of transmission of dengue virus. So the current study was planned to characterize the virus in Aedes mosquitoes collected from different area of Pakistan. In current investigation, Aedes mosquitoes and larvae were trapped under conducive conditions which are counted as 495 Aedes mosquitoes and 260 Aedes larvae. First of all, adult mosquitoes were identified morphologically under microscopy, counted as 73.3% Ae. aegypti and 26.7% Ae. albopictus. Finally, reverse transcriptase polymerase chain reaction analyses that only 4 adults of Aedes mosquitoes and 10 Aedes larvae as naturally infected with dengue virus with possible source Ae. aegypti. This study basically uncovered the presence of virus in different species of mosquitoes in southern regions of Pakistan. The present study will also give us an insight for vector control programs of dengue virus in the affected area.